Sweet pepper hybrid 9954288

ABSTRACT

The invention provides seed and plants of pepper varieties designated 9954288, SBR 99-1274 and SBY 99-1273. The invention thus relates to the plants, seeds and tissue cultures of sweet pepper hybrid 9954288, pepper line SBR 99-1274 and pepper line SBY 99-1273, and to methods for producing a pepper plant produced by crossing a plant with itself or with another pepper plant, such as a plant of another line. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of a plant of the invention, including the fruit and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Application Ser. No. 61/042,662, filed Apr. 4, 2008, the entire disclosure of which is incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, more specifically, to the development of sweet pepper hybrid 9954288, and of the inbred pepper lines SBR 99-1274 and SBY 99-1273.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include greater yield, resistance to insects or pests, tolerance to heat and drought, better agronomic quality, higher nutritional value, growth rate and fruit properties.

Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different varieties produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines are developed by selfing and selection of desired phenotypes. The new lines are evaluated to determine which of those have commercial potential.

One crop which has been subject to such breeding programs and is of particular value is the sweet pepper. As used herein, sweet pepper refers to the fruit and the plant of the non-pungent chile pepper varieties. Sweet peppers belong to the genre Capsicum, of the nightshade family, Solanaceae. The term “sweet pepper” therefore includes bell peppers (Capsicum annuum), the “That sweet”—also a cultivar of C. annuum, the “dulce”—a popular cultivar of Capsicum baccatum, as well as Numex Suave Orange (Capsicum chinense), an unusually sweet habanero-type pepper.

Sweet peppers are primarily used as vegetables in cuisine around the world; however, they are also grown for ornamental and medicinal uses. The sweet pepper fruit is eaten cooked or raw. In contrast to the hot pepper, the sweet pepper contains little, if any, capsaicin (8-methyl-N-vanillyl-6-nonenamide), the main active ingredient responsible for the pungency of hot peppers.

C. annuum is a herbaceous annual. The plant has a densely branched stem and grows to 1.5 to 5 feet in height. The fruit is green when unripe, then usually changing to red or brown. The species can grow in many climates; however, they thrive in warm and dry climates.

Most sweet pepper breeding programs have concentrated on the non-pungent varieties of C. annuum, especially bell peppers. Pickersgill and Barbara (1997). Breeding pepper lines with differently colored fruit has been very popular. The color of the fruit can be green, red, yellow, orange and, more rarely, white, purple and brown depending both on the cultivar and the time of harvest.

Peppers with multiple resistances to several pests and diseases have also been bred. Id. Pickersgill and Barbara (1997). In the case of bell peppers, the development of molecular markers and a molecular linkage map for C. annuum has eased some of the problems associated with selecting simultaneously for multiple resistances and other desirable characteristics. Id. Pickersgill and Barbara (1997). Other sweet pepper breeding efforts have often focused on creating non-pungent cultivars of hot peppers. See e.g. U.S. Pat. No. 7,087,819.

Tetraploidy and haploidy are relatively easy to induce in Capsicum species. In fact, an octaploid Capsicum annuum was recently reported (Panda et al., 1984). Capsicum species exhibit barriers to interspecific gene transfer. These include unilateral incompatibility, post-fertilization abortion, and nucleo-cytoplasmic interactions leading to male sterility or other abnormalities (Pickersgill and Barbara, 1997). However, the development of a pepper line resistant to the anthracnose fungal pathogen using interspecific crossing between Capsicum baccatum and C. annuum has been reported (Chae et al., 2003).

Hybrid vigor has also been documented in peppers, and hybrids are gaining increasing popularity among farmers throughout the world, especially in countries with plentiful labor (Berke, 1999).

While breeding efforts to date have provided a number of useful sweet pepper lines with beneficial traits, there remains a great need in the art for new lines with further improved traits. Such plants would benefit farmers and consumers alike by improving crop yields and/or quality.

SUMMARY OF THE INVENTION

The present invention overcomes limitations in the prior art by providing, for example, seeds and plants of a sweet pepper variety having a combination of genes, the expression of which provides a number of advantageous traits, such as small to medium plant size, and anthocyaninless, green to red bell pepper fruit with an large size at maturity of about 9 cm in width and 9 cm in length, and about 200 g to about 250 g. Other advantageous traits include resistance to: Race 6 Bacterial leaf spot (BLS) (Xanthomonas campestris pv. vesicatoria), Races 1-5 and 7-10 of BLS, Tobamovirus Pathotype 0 (TMV (p0)) and to Tomato spotted wilt virus (TSWV). This represents a significant advance in the art by offering resistance to Race 6 BLS, Races 1-5 BLS, Races 7-10 BLS and TSWV with the anthocyaninless trait, and excellent fruit qualities.

In one aspect, the present invention provides a pepper plant of the hybrid designated 9954288, and/or pepper lines SBR 99-1274 and SBY 99-1273. Also provided are pepper plants having all the physiological and morphological characteristics of the pepper hybrid 9954288. Parts of the sweet pepper plant of the present invention are also provided, for example, including pollen, an ovule, a scion, a rootstock, a fruit, and a cell of the plant.

In another aspect, the invention provides a plant of a sweet pepper hybrid that exhibits a combination of traits comprising resistance to: Race 6 BLS, Races 1-5 BLS, Races 7-10 BLS, TMV (p0), and TSWV, and anthocyaninless. In certain embodiments, the combination of traits may be defined as controlled by genetic means for the expression of the combination of traits found in sweet pepper hybrid 9954288.

The invention also concerns the seed of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. The pepper seeds of the invention may be provided as an essentially homogeneous population of pepper seed of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of pepper plants designated pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273.

In another aspect of the invention, a plant of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of the line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.

In another aspect of the invention, a tissue culture of regenerable cells of a pepper plant a plant described herein is provided. The tissue culture will preferably be capable of regenerating pepper plants capable of expressing all of the physiological and morphological characteristics of the line, and of regenerating plants having substantially the same genotype as other plants of the line. Examples of some of the physiological and morphological characteristics include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks. Still further, the present invention provides pepper plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of a plant provided herein.

In yet another aspect of the invention, processes are provided for producing pepper seeds, plants and fruit, which processes generally comprise crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. These processes may be further exemplified as processes for preparing hybrid pepper seed or plants, wherein a first pepper plant is crossed with a second pepper plant of a different, distinct line to provide a hybrid that has, as one of its parents, the pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent pepper plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first and second parent pepper plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the male portions of flowers, (i.e., treating or manipulating the flowers to produce an emasculated parent pepper plant). Self-incompatibility systems may also be used in some hybrid crops for the same purpose. Self-incompatible plants still shed viable pollen and can pollinate plants of other varieties but are incapable of pollinating themselves or other plants of the same line.

A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent pepper plants. Yet another step comprises harvesting the seeds from at least one of the parent pepper plants. The harvested seed can be grown to produce a pepper plant or hybrid pepper plant.

The present invention also provides the pepper seeds and plants produced by a process that comprises crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant provided herein such as from pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. In one embodiment of the invention, pepper seed and plants produced by the process are first generation (F₁) hybrid pepper seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F₁ hybrid pepper plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F1 hybrid pepper plant and seed thereof.

In still yet another aspect, the present invention provides a method of producing a plant derived from one or more of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid 9954288, wherein said preparing comprises crossing a plant of the pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273. The plant derived from pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 is obtained which possesses some of the desirable traits of the line as well as potentially other selected traits.

In certain embodiments, the present invention provides a method of producing peppers comprising: (a) obtaining a plant of pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273, wherein the plant has been cultivated to maturity, and (b) collecting peppers from the plant.

In still yet another aspect of the invention, the genetic complement of the pepper plant line designated pepper hybrid 9954288 and/or pepper lines SBR 99-1274 and SBY 99-1273 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a pepper plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides pepper plant cells that have a genetic complement in accordance with the pepper plant cells disclosed herein, and plants, seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that a plant provided herein could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

In still yet another aspect, the present invention provides hybrid genetic complements, as represented by pepper plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a pepper plant of the invention with a haploid genetic complement of a second pepper plant, preferably, another, distinct pepper plant. In another aspect, the present invention provides a pepper plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.

In still yet another aspect, the invention provides a method of determining the genotype of a plant described herein comprising detecting in the genome of the plant at least a first polymorphism. The method may, in certain embodiments, comprise detecting a plurality of polymorphisms in the genome of the plant. The method may further comprise storing the results of the step of detecting the plurality of polymorphisms on a computer readable medium. The invention further provides a computer readable medium produced by such a method.

Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.

The term “about” is used to indicate that a value includes the standard deviation of error for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and to “and/or.” When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.

Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants, seeds and derivatives of sweet pepper hybrid 9954288, pepper line SBY 99-1273 and pepper line SBR 99-1274. Hybrid 9954288 can be described as a green to red bell pepper variety. The traits of this variety include a small to medium plant size, golden yellow anthers and anthocyaninless. Anthocyanin is not observed in either the fruit or the stem. The mature fruit of the hybrid 9954288 can be further characterized as being firm and having a weight from about 200 to about 250 grams, a size that is large to extra large, and a typically blocky bell pepper shape. Average fruit dimensions at maturity are 8-10 cm wide by 8-10 cm long. Fruit at the green stage is dark green, while fruit at red stage is bright red.

Sweet pepper hybrid 9954288 exhibits resistance to: Race 6 BLS (Xanthomonas campestris pv. vesicatoria), Races 1-5 and Races 7-10 of BLS, TMV (p0) and TSUV. This variety shows uniformity and stability within the limits of environmental influence for the traits described hereinafter. Sweet pepper hybrid 9954288 provides sufficient seed yield.

Sweet pepper hybrid 9954288 exhibits a number of improved traits including resistance to: Race 6 of BLS, Race 1-5 BLS, Race 7-10 BLS, TMV (p0), and to TSWV. A Race 6 resistant, Race 1-5 resistant, and Race 7-10 resistant hybrid will allow growers to continue to grow and harvest peppers in areas affected by Race 6, Races 1-5 and Races 7-10 BLS, TMV (p0), and the TSWV, for example in open-field commercial production in places, such as in Georgia, USA, including the southern parts of that state. Without this resistance it is often impossible to prevent heavy infection in areas where Race 6, Race 1-5, Race 7-10 BLS is present and environmental conditions favor BLS infection. Other suitable places of production and sale are expected to include, for example, Venezuela, South Africa, Thailand, Australia, parts of the Caribbean, and Italy. The development of the variety can be summarized as follows.

A. ORIGIN AND BREEDING HISTORY OF SWEET PEPPER HYBRID 9954288

The parents of sweet pepper hybrid 9954288 are pepper line SBR 99-1274 and pepper line SBY 99-1273. These parents were both created at breeding stations in Felda, Fla. and San Juan Bautista, Calif. Race 6 resistance was originally provided by line 4413-23-1BK (Robert Stall, University of Florida, Gainesville). TSWV resistance was provided from Seminis bell pepper parent line YTSWV9796-2.

Inbred SBR 99-1274 was created by crossing “F2RNS/1091.PC01-1101-5,” a red, blocky bell segregating for Race 6 BLS resistance ((Xanthomonas campestris pv. vesicatoria) bs5 and bs6 genes) to YTSWV9796-2, which is a yellow, blocky bell fixed for resistance to TSWV. Neither parent was marketed as open pollinated lines. SBR 99-1274 is fixed for resistance to race 6 BLS, is resistant to the TSWV, and is red. Thus, SBR 99-1274 differs from both of its parents, neither of which has this combination of characteristics. The crossing and selections that led to the development of SBR 99-1274 can be summarized as follows:

Year 1 Planted parents “F2RNS/1091.PC01-1101-5” and “YTSWV9796-2” in greenhouses at San Juan Bautista Research Station. The F₁ Hybrid was made.

Year 2 Planted F₁ hybrid SE-PC01-1101-5×SE-PC01-1046 as stake PC02 03046. Individual plants were selfed.

Year 2 Planted F₂ inbred PC02 03046-9 as stake PC02 FL 1273. Individual plant selection made.

Year 3 Planted F₃ inbred PC02 FL 1273-3 as stake 03LB 00417. Fruit set, shape and size were adequate. Plants were segregating for bacterial spot Race 6 (Xanthomonas campestris pv. vesicatoria) and TSWV. Plant selections were made.

Year 3 Planted F₄ inbred 03LB 00417-01 as stake 03LB 06282. Plants produced very large, red, hourglass shaped fruits. The line tested susceptible for bacterial spot Races 1 to 5 (Bs1, Bs2, Bs3 genes) but resistant to Race 6. The line was segregating for TSWV. The line was bulked. Year 4 Planted F₅ inbred line of 03LB 06282-M as stake 04LB 00405. The line was fixed for the anthocyaninless trait. The line tested fixed for resistance to the TSWV. Plant selections were made. Year 4 Planted F₆ inbred line 04LB 00405-4 as stake 04LB 05541. The plants produced a heavy set of red, broad-shouldered fruit, with adequate firmness and green color. Plants tested resistant to Tobacco mosaic virus (L1 gene) and resistant to Race 6 (Xanthomonas campestris pv. vesicatoria). The line tested fixed for resistance to the TSWV. Plant selections were made. Year 5 Planted F₇ inbred line 04LB 05541-03 as stake 05LB LBGH 0197. Line appeared uniform and stable. The line was bulked. Year 5 Planted F₈ inbred line bulk 05LB LBGH 0197-M as stake 05LB 06526 for increase. Line produced large to extra large red fruit of adequate color, heavy to adequate set, with adequate firmness. Observations during the growing season indicted the line was uniform and stable. The entire plot was selected and bulked. Year 6 Planted F₉ inbred line bulk 05LB 06526-M as 06LB LBGH 1454-M for increase.

SBR 99-1274 is uniform and stable. It is within commercial acceptable limits as is true with other Sweet Pepper inbred lines. A small percentage of variants can occur within commercially acceptable limits for almost any character during the course of repeated multiplication. However no variants were observed, during the four times in which SBR99-1274 was observed in other trials.

SBY 99-1273 was developed by a cross between PC01-1089-8-F2RNSXPPR, a bell pepper with blocky yellow fruit segregating for Race 6 bacterial spot resistance (Xanthomonas campestris pv. vesicatoria) through bs5 and bs6 genes, and SBR27-1004, which is a bell pepper with blocky red fruit fixed for bacterial spot resistance (Xanthomonas campestris pv. vesicatoria) through Bs2 gene, and fixed for TMV (p0) (L1 gene), and fixed for Potato virus Y pathotype P₀ (pvr2-2 gene). Neither parent was marketed as an open pollinated line. SBR 99-1273 is fixed for resistance to Races 3 and 6 bacterial spot (Xanthomonas campestris pv. vesicatoria) and carries Bs2, bs5 and bs6 genes, is fixed for resistance to the TMV (p0) (L1 gene), and has yellow fruit.

The crossing and selections that led to the development of line SBY 99-1273 can be summarized as follows:

Year 1 Parents planted, PC01-1089-8-F2RNSXPPR as stake PC01 1089-8 and SBR27-1004 as stake PC01 10522 in greenhouses at San Juan Bautista Research Station, Calif. The F₁ Hybrid was made.

Year 2 F₁ hybrid planted, PC02 03017 at San Juan Bautista Research Station. Individual plants were selfed.

Year 2 F₂ inbred line planted of PC02 03017-6 at the Felda Research Station in Florida as stake PC02FL 01 194. Plant selections were made.

Year 3 F₃ inbred line was planted of PC02FL 01194-03 as stake 03LB 00204 at the Felda Research Station in Florida. Plants were segregating for resistance for bacterial spot Race 6 (Xanthomonas campestris pv. vesicatoria). The line was found susceptible to Phytophthora capsici. Plant selections were made. Year 3 F₄ inbred line was planted of 03LB 00204-01 as stake 03LB 06407. Line was found homozygous for anthocyaninless. Pathology tests indicate the line was fixed resistant to Races 1, 2 and 3 of bacterial spot ((Xanthomonas campestris pv. vesicatoria) Bs2 gene) and segregating for resistance to Race 6 (bs5 bs6 genes). Plant selections were made. Year 4 F₅ inbred line was planted of 03LB 06407-01 as stake 04LB 00589. The line was found fixed for yellow mature fruits. Pathology tests indicate the line was segregating for resistance to Race 6 (Xanthomonas campestris pv. vesicatoria). A single plant selection was made. Year 4 F₆ inbred line was planted of 04LB 00589-1 as stake 04LB 05652. Pathology tests indicate the line was fixed for resistance to TMV (p0) (L1 gene) and segregating for resistance to Race 6 (Xanthomonas campestris pv. vesicatoria). Plant selections were made. Year 5 F₇ inbred line was planted, 04LB 05652-01 as stake 05LB 00529. Line was fixed for upright peduncle. Line was fixed for resistance to bacterial spot Race 6 (Xanthomonas campestris pv. vesicatoria). The line was observed to be uniform and stable. The line was bulked. Year 5 F₈ bulk inbred line was planted, 05LB 00529-M as stake 05LB 06559. The plants were small and produced slightly flat fruit that were very firm and small to medium in size. The line was resistant to bacterial spot Race 6 (Xanthomonas campestris pv. vesicatoria). The line was found to be stable and uniform. The line was bulked. Year 6 F₉ bulk inbred line was planted, 05LB 06559-M as stake 06LB 03152. The line was found to produce a heavy fruit set of very firm fruit, small to medium in size with four lobes. Fruit were medium green in color and the plant size was small. The line was found resistant to bacterial spot ((Xanthomonas campestris pv. vesicatoria) Bs2 and bs5 bs6 genes). The line was bulked.

SBR 99-1273 is uniform and stable. It is within commercially acceptable limits as is true with other Sweet Pepper inbreds. A small percentage of variants can occur within commercially acceptable limits for almost any character during the course of repeated multiplication. However no variants were observed, during the two times in which SBR99-1273 was observed in other trials.

B. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF PEPPER HYBRID 9954288

In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of sweet pepper hybrid 9954288. A description of the physiological and morphological characteristics of sweet pepper hybrid 9954288 is presented in Table 1.

TABLE 1 Physiological and Morphological Characteristics of Pepper Hybrid 9954288 CHARACTERISTIC HYBRID 9954288 Species C. annuum Mature Plant Days from Transplanting until 75 Green Stage Plant Habit Semi-Spreading Plant Attitude Semi-Upright/Semi-Erect Stem: Hairiness of Nodes Absent or Very Weak Height Medium Basal Branches Few (2-3) Branch Flexibility Rigid Leaf Length of Blade Medium Width of Blade Medium Color Medium Green Intensity of Green Color Medium Mature Leaf Shape Broad Elliptic Leaf and Stem Pubescence Light Undulation of Margin Weak Profile in Cross Section Flat Peduncle Attitude Drooping Flower Corolla Color White Anther Color Yellow Self-Incompatibility Absent Fruit Group Bell (Yolo Wonder L.) Color (Before Maturity) Green Intensity of Color (Before Medium Maturity) Color (Immature Fruit) Medium Attitude/Position Drooping/Pendent Length Medium Diameter Broad Ratio Length/Diameter Medium Diameter at Mid-Point 1000.0 mm Flesh Thickness at Mid-Point 7.0 mm Average Number of Fruits per 10 Plant Average Weight 220 gm Shape in Longitudinal Section Square Shape in Cross-Section (Level Quadrangular of Placenta) Texture of Surface Smooth/Very Slightly Wrinkled Color (Mature) Red Intensity of Color at Maturity Medium Glossiness Medium Stalk Cavity Present Depth of Stalk Cavity Medium Stalk Length Medium Stalk Thickness Medium Base Shape Cupped Shape of Apex Moderately Depressed Shape Bell (Yolo Wonder L.) Set Concentrated Depth of Interloculary Grooves Shallow Number of Locules Predominately four and More Measurement with Three 50% Locules Measurement with Four 40% Locules Measurement with Five or 10% More Thickness of Flesh Thick Calyx: Aspect Non-Enveloping/Saucer-Shaped Pungency Sweet Capsaicin in Placenta Absent Flavor - Dry Fruit Mild Pepper Glossiness - Dry Fruit Moderate Seed Color Yellow Anthocyanin Coloration of Absent Hypocotyl Plant Anthocyanin Coloration of Absent Stem Anthocyanin Coloration of Absent Nodes Stem: Intensity of Very Weak Anthocyanin Coloration of Nodes Flower: Anthocyanin Absent Coloration in Anther Fruit: Anthocyanin Coloration Absent Beginning of Flowering (1^(st) Medium Flower on 2^(nd) Flowering Node) Time of Maturity Medium Diseases Resistance to Tobamovirus Most Resistant Pathotype 0 Resistance to Tomato Spotted Most Resistant Wilt virus (TSWV) Resistance to Xanthomonas Most Resistant campestris pv. vesicatoria Resistance to Nematode Most Susceptible (Meloidogyne incognita acrita) Resistance to Ripe Rot Most Susceptible (Vermicularia capsici) Resistance to Southern Blight Most Susceptible (Sclerotium rolfsii) Resistance to Verticillium Wilt Most Susceptible (Verticillium dahliae) Resistance to Race 6 Most Resistant Xanthomonas campestris pv. vesicatoria *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are within the scope of the invention.

The parents of hybrid 9954288, SBR99-1274 and SBY 99-1273, have been self-pollinated and planted for a number of generations to produce the homozygosity to make the hybrid 9954288 phenotypically stable and commercial useful. No variant traits have been observed or are expected for the parents of this hybrid.

C. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF BREEDING SWEET PEPPER SBR99-1274

The hybrid 9954288 was produced by crossing inbred SBR 99-1274 with SBY 99-1273. A description of the physiological and morphological characteristics of pepper lines SBR 99-1274 and SBY 99-1273 is presented in Tables 2 and 3.

TABLE 2 Physiological and Morphological Characteristics of Pepper SBR 99-1274 CHARACTERISTIC SBR 99-1274 Species C. annuum Mature Plant Days from Transplanting until 75 Green Stage Maturity (in region of best 90 adaptability) Plant Habit spreading Color Medium Green Intensity of Green Color Medium (Doux tres long des Landes, Merit) Mature Leaf Shape Broad Elliptic Peduncle Attitude Drooping Fruit Group Bell (Yolo Wonder L.) Color (Before Maturity) Green (California wonder, Lamuyo) Intensity of Color (Before Medium Maturity) Intensity of Color (Before Medium Maturity) Color (Immature Fruit) Dark Green Attitude/Position Drooping/Pendent (De Cauyenne, Lamuyo) Length Medium (Feher, Lamuyo) Diameter Broad (Clovis, Lamuyo) Ratio Length/Diameter Medium (Adra, Cherry Sweet, Daniel, Delphin, Edino) Fruit Measurements: Fruit 1100.0 mm length Diameter at Mid-Point 1100.0 mm Shape in Longitudinal Section Square (Delphin, Yolo Wonder) Shape in Cross-Section (Level Quadrangular of Placenta) Texture of Surface Smooth/Very Slightly Wrinkled (Milord) Intensity of Color at Maturity Medium Mature fruit color Red Shape of Apex Very depressed (Kerala, Monte, Osir) Shape Bell (Yolo Wonder L.) Depth of Interloculary Grooves Medium (Clovis, Lamuyo, Marconi) Thickness of Flesh Thick (Andevalo, Bingor, Daniel, Topgirl) Pungency Sweet Capsaicin in Placenta Absent Flavor - Dry Fruit Mild Pepper Flavor Seed Color Yellow Anthocyanin Coloration of Absent (Albaregia, Albena) Hypocotyl Plant Anthocyanin Coloration of Absent Stem Anthocyanin Coloration of Absent Nodes Flower: Anthocyanin Absent Coloration in Anther Fruit: Anthocyanin Coloration Absent Time of Maturity Late (Daniel, Doux d'Espagne) Diseases Resistance to Tobamovirus Most Resistant (Lamuyo, Sonar, Yolo Wonder) Pathotype 0 (Tobacco Mosaic Virus (0)) Resistance to Tomato Spotted Present (Galileo, Jackal, Jackpot) Wilt virus (TSWV) Resistance to Xanthomonas Present/Most Resistant (Aladin, campestris pv. vesicatoria Camelot, EOR-20R, Kaldom, Kalorex, Lancelot, Pasa) Resistance to Race 4-10 of Most Resistant bacterial spot *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are within the scope of the invention.

TABLE 3 Physiological and Morphological Characteristics of Pepper SBY 99-1273 and a comparison variety CHARACTERISTIC SBY 99-1273 Early Cal Wonder Species C. annuum C. annuum Maturity (in region of best adaptability) Days from transplanting 77 67 until mature green stage Days from transplanting 100 85 until mature red or yellow stage Days from direct seeding 114 104 until mature green stage Days from direct seeding 137 122 until mature red or yellow stage Plant Habit Compact Compact Attitude Upright/erect (De Upright/erect Cayenne, Doux très long des Landes, Piquant d'Algérie) Plant height 35.4 cm 40.9 cm Plant width 38 cm 47.1 cm Length of stem from 14 cm 10.7 cm cotyledon to first flower Length of the third internode 60 mm 54.0 mm (from soil surface) Length of stem Short (Delphin, Trophy) Shortened internode (in Absent (California upper part) wonder, De Cayenne) For varieties with shortened None (Kalocsai 601) internodes only: Plant: number of internodes between the first flower and the shortened internodes For varieties with shortened Medium (Dolmi, internodes only: Plant: Florian, Órias) length of internode (on primary side shoots) Stem: hairiness of nodes Absent or very weak (Arlequin) Height Short (Albaregia) Basal branches None Few Branch flexibility Rigid (Yolo Wonder) Rigid Stem strength (breakage Strong Intermediate resistance) Length of blade Long (Cupido, Dolmy, Encore, Mazurka, Monte) Width of blade Broad (California wonder, Golden calwonder, Seifor, Solario) Leaf Leaf width 11.7 mm 11.3 mm Leaf length 6.6 mm 6 mm Petiole length 4.6 mm 4.6 mm Color Light green Light green RHS Color Chart Value 146A 147A Intensity of green color Medium (Doux très long des Landes, Merit) Mature leaf shape Ovate (Balico, Sonar) Ovate Leaf and stem pubescence Moderate Absent Undulation of margin Absent (De Cayenne) Absent Blistering Absent (Century, Weak Recio, Solfiane) Profile in cross section Moderately concave (doux italien, Favolor) Glossiness Strong (Andevalo, Floridor) Peduncle: attitude Erect (Fehér, Red Chili) Flower Flowers per leaf axil 1 1 Calyx lobes 6 6 Petals 6 6 Diameter 2.69 mm 2.2 mm Corolla color White White Corolla throat markings Yellow Yellow Anther color Yellow Purple Style length Less than stamen Same as stamen Self-incompatibility Absent Absent Fruit Group Bell (Yolo Wonder L.) Bell (Yolo Wonder L.) Color (before maturity) Green (California wonder, Lamuyo) Intensity of color (before Medium maturity) Immature fruit color Medium green Medium green Immature fruit color 146A 137A RHS Color Chart value Attitude/position Erect/upright Drooping/pendant (Kalocsai 601, Red Chili) Length Medium (Fehér, Lamuyo) Diameter Broad (Clovis, Lemuyo) Ratio length/diameter Small (Bucano, Topgirl) Calyx diameter 29.25 mm 32.0 mm Length 68.25 mm 80.0 mm Diameter at calyx 63.1 mm 70.0 mm attachment Diameter at mid-point 70 mm 80.0 mm Flesh thickness at mid-point 5.2 mm 6.0 mm Average number of fruits 9.4 10.0 per plant % large fruits (weight range: (weight range: 141 to 200 g) 130 to 200 g) 6.03% 50.0% % medium fruits (weight range: (weight range: 81 to 140 g) 90 to 120 g) 33.9% 30.0% % small fruits (weight range: (weight range: 20 to 80 g) 50 to 75 g) 60.07% 20.0% Average fruit weight 96.5 gm 100.0 gm Shape in longitudinal Square (Delphin, Yolo Square section Wonder) Shape in cross section (at Quadrangular Quadrangular level of placenta) Sinuation of pericarp at Weak (Donat) basal part Sinuation of pericarp Absent or very weak excluding basal part (Delphin, Milord) Texture of surface Smooth or very slightly Smooth or very wrinkled (Milord) slightly wrinkled Color (at maturity) Yellow (Golden calwonder, Heldor) Intensity of color (at Medium maturity) Mature fruit color Orange-yellow Red Mature fruit color RHS 16A 46A Color Chart value Glossiness Medium/moderate Medium/moderate (Carré doux extra hâtif, Lamuyo, Sonar) Pedicel length 20.0 mm Pedicel thickness 8.6 mm 6.0 mm Pedicel shape Straight Curved Pedicel cavity Present Absent Stalk: length Medium (Fehér, Sonar) Stalk: thickness Thick (Lamuyo, Trophy Palio) Base shape Cupped Cupped Shape of apex Rounded (Cherry Very depressed Sweet) Shape Bell (Yolo Wonder L.) Bell (Yolo Wonder L.) Set Concentrated Scattered Depth of interloculary Medium (Clovis, Medium grooves Lamuyo, Marconi) % fruits with one locule 0% 0% % fruits with two locules 3.03% 0% % fruits with three locules 60.6% 40.0% % fruits with four locules 33.34% 60.0% % fruits with five or more 3.03% 0% locules Average number of locules 3.36 3.6 Thickness of flesh Medium ((Fehér, Lamuyo) Calyx: aspect Non-enveloping/ Non-enveloping/ saucer-shaped saucer-shaped (Lamuyo, Sonar) Pungency Sweet Sweet Capsaicin in placenta Absent (Sonar) Flavor Moderate Glossiness Shiny Shiny Seed Seed cavity length 49.15 mm 43.0 mm Seed cavity diameter 59.6 mm 52.0 mm Placenta length 21.15 mm 22.0 mm Number of seeds per fruit 98 100 Grams per 1000 seeds 7.5 gm Color Yellow Seedling Anthocyanin coloration of Absent (Albaregia, Moderate hypocotyl Albena) Plant: anthocyanin Absent Absent coloration of stem Plant: anthocyanin Weak Weak coloration of nodes Stem: intensity of Very weak anthocyanin coloration of nodes Plant: anthocyanin Absent Absent coloration of leaf Plant: anthocyanin Absent Absent coloration of pedicel Plant: anthocyanin Absent Absent coloration of calyx Flower: anthocyanin Absent (Danza) coloration in anther Fruit: anthocyanin Absent (Lamuyo) Absent coloration Beginning of flowering (1^(st) Medium (Lamuyo, flower on 2^(nd) flowering Latino) node) Time of maturity Medium (Lamuyo, Latino, Sonar) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are within the scope of the invention.

D. BREEDING SWEET PEPPER HYBRID 9954288 AND PARENTS THEREOF

One aspect of the current invention concerns methods for crossing the sweet pepper hybrid 9954288 and parent lines thereof with itself or a second plant and the seeds and plants produced by such methods. Further hybrid variety seeds are produced by crossing sweet pepper hybrid 9954288 with second pepper parent line or variety

The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing sweet pepper hybrid 9954288 and/or parent lines thereof followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines or varieties, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with sweet pepper hybrid 9954288 and parent lines or progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny are heterozygous for loci controlling the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.

The line of the present invention is particularly well suited for the development of new lines based on the elite nature of the genetic background of the line. In selecting a second plant to cross with sweet pepper hybrid 9954288 or a parent thereof for the purpose of developing novel sweet pepper lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits of sweet peppers include: high seed yield, high seed germination, seedling vigor, early fruit maturity, high fruit yield, ease of fruit setting, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a preference for a given fruit size, shape, color, texture, and taste, especially non-pungency (low capsaicinoid content), are other traits that may be incorporated into new lines of sweet pepper plants developed by this invention.

Particularly desirable traits that may be incorporated by this invention is improved resistance to different viral, fungal, and bacterial pathogens. Anthracnose and Phytophthora blight are fungal diseases affecting various species of pepper. Fruit lesions and fruit rot are the commercially important aspects of these diseases. Bacterial leaf spot and bacterial wilt are other diseases affecting pepper plants, especially during the wet season. Viral pathogens affecting pepper plants include the Pepper mosaic virus and the Tobacco mosaic virus.

Improved resistance to insect pests is another desirable trait that may be incorporated into new lines of pepper plants developed by this invention. Insect pests affecting the various species of pepper include the European corn borer, corn earworm, aphids, flea beetles, whiteflies, and mites (Midwest Vegetable Production Guide for Commercial Growers, 2003).

E. PERFORMANCE CHARACTERISTICS OF HYBRID 9954288

As described above, hybrid 9954288 exhibits desirable agronomic traits, including anthocyaninless, a fruit size at maturity from about 200 g to about 300 g and resistance to: Race 6 BLS (Xanthomonas campestris pv. vesicatoria), Races 1-5 BLS and Races 7-10 BLS, TMV (p0), and to TSWV. A non-hypersensitive resistance in pepper to the bacterial spot pathogen is associated with two recessive genes (Jones and Minisavage, 2002).

While there is an open-pollinated pepper inbred available with Race 6 resistance called ECW12346, it has much smaller fruit size, is susceptible to TSWV, and is not anthocyaninless. There is another open-pollinated pepper inbred available with moderate tolerance to Race 6 called “Dempsey”, but it is not anthocyaninless, nor does it have TSWV resistance (see Lane et al., 1997)

There are multiple commercial hybrids available with TSWV resistance but none having Race 6 BLS resistance. These and other performance characteristics of the line were the subject of an objective analysis of the performance traits of the line relative to other lines. The results of the analysis are presented below.

TABLE 3 Performance Characteristics For Hybrid 9954288 Moderate Fruit fruit size Race 1-3 Race 6 Phytophthora color at Width (cm) × Fruit BLS BLS TSWV TMV (Po) capsici antho- green length weight Variety source resistance resistance resistance resistance resistance cyaninless harvest (cm) (g.) 9954288 Seminis yes yes yes yes No no medium Aristotle Seminis yes no no yes yes yes medium 10 × 10 300 Plato Seminis yes no yes yes no no medium- 10 × 12 300 dark Stiletto Syngenta yes no yes no no yes medium 8 × 8 200 Polaris Western yes no no yes no no dark 10 × 10 300 Seeds Dempsey Syngenta no tolerance no yes no no medium 6 × 6 150

F. FURTHER EMBODIMENTS OF THE INVENTION

In one embodiment, the invention provides plants of pepper hybrid 9954288 and parent lines thereof modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those pepper plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique.

Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental pepper plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental pepper plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a pepper plant is obtained wherein essentially all of the desired morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.

In one embodiment, progeny pepper plants of a backcross in which a parent of 9954288 is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of the parent of sweet pepper hybrid 9954288 as determined at the 5% significance level when grown in the same environmental conditions.

Pepper varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.

Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, male sterility, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, restoration of male fertility, modified fatty acid or carbohydrate metabolism, and enhanced nutritional quality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as a dominant trait. An example of a dominant trait is the downy mildew resistance trait. For this selection process, the progeny of the initial cross are sprayed with downy mildew spores prior to the backcrossing. The spraying eliminates any plants which do not have the desired downy mildew resistance characteristic, and only those plants which have the downy mildew resistance gene are used in the subsequent backcross. This process is then repeated for all additional backcross generations.

Selection of pepper plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection applicable to the breeding of pepper are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).

G. PLANTS DERIVED FROM SWEET PEPPER HYBRID 9954288 AND PARENT LINES THEREOF BY GENETIC ENGINEERING

Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into the pepper line of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants, including pepper plants, are well known to those of skill in the art (see, e.g., below). Techniques which may be employed for the genetic transformation of pepper plants include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.

Agrobacterium-mediated transformation of pepper explant material and regeneration of whole transformed pepper plants (including tetraploids) from the transformed shoots has been shown to be an efficient transformation method (U.S. Pat. No. 5,262,316).

A particularly efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target pepper cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. It is believed that a screen intervening between the projectile apparatus and the cells to be bombarded reduces the size of projectiles aggregate and may contribute to a higher frequency of transformation by reducing the damage inflicted on the recipient cells by projectiles that are too large.

Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986; Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (1994), and Ellul et al. (2003).

A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for pepper plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., 1985), including monocots (see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S) the nopaline synthase promoter (An et al., 1988), the octopine synthase promoter (Fromm et al., 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem, the cauliflower mosaic virus 19S promoter, a sugarcane bacilliform virus promoter, a commelina yellow mottle virus promoter, and other plant DNA virus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter, Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos et al., 1989).

Exemplary nucleic acids which may be introduced to the pepper lines of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and could potentially be introduced into a pepper plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a pepper plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880275, herein incorporated by reference it their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.

H. DEFINITIONS

In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F₁), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes from different plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenic plants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent. Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the desired morphological and physiological characteristics of a pepper variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.

Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.

Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.

I. DEPOSIT INFORMATION

A deposit of sweet pepper hybrid 9954288, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit was Oct. 16, 2007. The accession number for those deposited seeds of sweet pepper hybrid 9954288 is ATCC Accession No. PTA-8690. A deposit of sweet pepper SBR 99-1274, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit was Oct. 16, 2007. The accession number for those deposited seeds of sweet pepper SBR 99-1274, is ATCC Accession No. PTA-8689. A deposit of sweet pepper SBY 99-1273, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit was May 30, 2008. The accession number for those deposited seeds of sweet pepper SBY 99-1273, is ATCC Accession No. PTA-9229.

Upon issuance of a patent, all restrictions upon the foregoing deposits will be removed. The deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.

All references cited herein are hereby expressly incorporated herein by reference.

REFERENCES

The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference:

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1. A pepper plant comprising at least a first set of the chromosomes of pepper line SBR 99-1274 or pepper line SBY 99-1273, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively.
 2. A seed comprising at least a first set of the chromosomes of pepper line SBR 99-1274 or pepper line SBY 99-1273, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively.
 3. The plant of claim 1, which is inbred.
 4. The plant of claim 1, which is hybrid.
 5. The plant of claim 4, wherein the hybrid plant is pepper hybrid 9954288, a sample of seed of said hybrid 9954288 having been deposited under ATCC Accession Number PTA-8690.
 6. The plant of claim 3, wherein the inbred plant is a plant of line SBR 99-1274 or line SBY 99-1273.
 7. A plant part of the plant of claim
 1. 8. The plant part of claim 7, further defined as a leaf, a ovule, pollen, a fruit, or a cell.
 9. A pepper plant, or a part thereof, having all the physiological and morphological characteristics of the pepper plant of claim
 5. 10. A pepper plant, or a part thereof, having all the physiological and morphological characteristics of the pepper plant of claim
 6. 11. A tissue culture of regenerable cells of the plant of claim
 1. 12. The tissue culture according to claim 11, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
 13. A pepper plant regenerated from the tissue culture of claim 12, wherein the plant has all the physiological and morphological characteristics of a pepper plant comprising at least a first set of the chromosomes of pepper line SBR 99-1274 or pepper line SBY 99-1273, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively.
 14. A method of vegetatively propagating the plant of claim 1 comprising the steps of: (a) collecting tissue capable of being propagated from a plant according to claim 1; (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
 15. The method of claim 14, further comprising growing at least a first plant from said rooted plantlets.
 16. A method of introducing a desired trait into a pepper line comprising: (a) utilizing as a recurrent parent a plant of either pepper line SBR 99-1274 or pepper line SBY 99-1273, by crossing a plant of line SBR 99-1274 or line SBY 99-1273, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively, with a second donor pepper plant that comprises a desired trait, to produce F₁ progeny; (b) selecting an F₁ progeny that comprises the desired trait; (c) crossing the selected F₁ progeny with a plant of the same pepper line used as the recurrent parent in step (a) [line SBR 99-1274 or SBY 99-1273] to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristics of the recurrent parent pepper line used in step (a) [pepper line SBR 99-1274 or line SBY 99-1273]; and (e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait, and otherwise comprise essentially all of the physiological and morphological characteristics of the recurrent parent pepper line used in step (a).
 17. A pepper plant produced by the method of claim
 16. 18. A method of producing a plant comprising an added trait, the method comprising introducing a transgene conferring the trait into a plant of hybrid 9954288, line SBR 99-1274 or line SBY 99-1273, a sample of seed of said hybrid and lines having been deposited under ATCC Accession Number PTA-8690, ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively.
 19. A plant produced by the method of claim
 18. 20. A method of determining the genotype of the plant of claim 1 comprising obtaining a sample of nucleic acids from said plant and detecting in said nucleic acids a plurality of polymorphisms.
 21. The method of claim 20, further comprising the step of storing the results of detecting the plurality of polymorphisms on a computer readable medium.
 22. A method for producing a seed of a plant derived from at least one of hybrid 9954288, line SBR 99-1274 or line SBY 99-1273 comprising the steps of: (a) crossing a pepper plant of hybrid 9954288, line SBR 99-1274 or line SBY 99-1273 with itself or a second pepper plant; a sample of seed of said hybrid and lines having been deposited under ATCC Accession Number PTA-8690, ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively; and (b) allowing seed of a hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper plant to form.
 23. The method of claim 22, further comprising the steps of: (c) selfing a plant grown from said hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper seed to yield additional hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper seed; (d) growing said additional hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper seed of step (c) to yield additional hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper plant.
 24. The method of claim 22, wherein the second pepper plant is of an inbred pepper line.
 25. The method of claim 22, comprising crossing line SBR 99-1274 with line SBY 99-1273, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-8689, and ATCC Accession Number PTA-9229, respectively.
 26. The method of claim 23, further comprising: (f) crossing the further hybrid 9954288, line SBR 99-1274 or line SBY 99-1273-derived pepper plant with a second pepper plant to produce seed of a hybrid progeny plant.
 27. A hybrid seed produced by the method of claim
 25. 28. A plant produced by growing the seed of claim
 27. 29. A plant part of the plant of claim
 28. 30. The plant part of claim 29, further defined as a leaf, a flower, a fruit, an ovule, pollen, or a cell.
 31. The seed of claim 27, wherein one or both of the plant of line SBR 99-1274 or line SBY 99-1273 and the second plant comprises a transgene.
 32. The seed of claim 27, wherein one or both of the plant of line SBR 99-1274 or line SBY 99-1273 and the second plant comprises a single locus conversion.
 33. A method of producing a pepper seed comprising crossing the plant of claim 1 with itself or a second pepper plant and allowing seed to form.
 34. A method of producing a pepper fruit comprising: (a) obtaining a plant according to claim 1, wherein the plant has been cultivated to maturity; and (b) collecting a pepper from the plant.
 35. The method of claim 34, wherein the plant according to claim 1 is a plant of pepper hybrid 9954288, a sample of seed of said hybrid 9954288 having been deposited under ATCC Accession Number PTA-8690. 